Anti-TDRD9 antibody (ab118427)
Key features and details
- Rabbit polyclonal to TDRD9
- Suitable for: WB, ICC/IF, IHC-P
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Overview
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Product name
Anti-TDRD9 antibody -
Description
Rabbit polyclonal to TDRD9 -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IF, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Chimpanzee, Macaque monkey, Gorilla
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Mouse and rat testis tissue lysate. IHC-P: Human normal testis tissue. ICC/IF: Hek293 cells.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab118427 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application Abreviews Notes WB Use a concentration of 1 µg/ml. Detects a band of approximately 171 kDa (predicted molecular weight: 155 kDa). Abcam recommends using milk as the blocking agent (3%). ICC/IF Use a concentration of 5 µg/ml. IHC-P Use a concentration of 5 µg/ml. Target
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Function
Probable ATP-binding RNA helicase which plays a central role during spermatogenesis by repressing transposable elements and prevent their mobilization, which is essential for the germline integrity. Acts via the piRNA metabolic process, which mediates the repression of transposable elements during meiosis by forming complexes composed of piRNAs and Piwi proteins and govern the methylation and subsequent repression of transposons. Its association with PIWIL4 and the piP-bodies suggests a participation in the secondary piRNAs metabolic process. -
Sequence similarities
Belongs to the DEAD box helicase family. DEAH subfamily.
Contains 1 helicase ATP-binding domain.
Contains 1 helicase C-terminal domain.
Contains 1 Tudor domain. -
Cellular localization
Cytoplasm. Nucleus. Component of the nuage, also named P granule, a germ-cell-specific organelle required to repress transposon during meiosis. Specifically localizes to piP-bodies, a subset of the nuage which contains secondary piRNAs. PIWIL2 is required for its localization to piP-bodies. - Information by UniProt
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Database links
- Entrez Gene: 122402 Human
- Entrez Gene: 74691 Mouse
- Entrez Gene: 299343 Rat
- SwissProt: Q8NDG6 Human
- SwissProt: Q14BI7 Mouse
- SwissProt: Q3MHU3 Rat
- Unigene: 21454 Human
- Unigene: 60648 Mouse
see all -
Alternative names
- C14orf75 antibody
- chromosome 14 open reading frame 75 antibody
- DKFZp434N0820 antibody
see all
Images
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Western blot - Anti-TDRD9 antibody (ab118427)Anti-TDRD9 antibody (ab118427) at 1 µg/ml (Milk blocking - 3%) + Testis (Mouse) Tissue Lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 155 kDa
Observed band size: 171 kDa why is the actual band size different from the predicted?
Additional bands at: 190 kDa, 50 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 8 minutes -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TDRD9 antibody (ab118427)IHC image of TDRD9 staining in Human normal testis formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab118427, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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Western blot - Anti-TDRD9 antibody (ab118427)All lanes : Anti-TDRD9 antibody (ab118427) at 1 µg/ml
Lane 1 : MCF7 cells (human).
Lane 2 : MDA231 cells (human).
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 155 kDa
Observed band size: 170 kDa why is the actual band size different from the predicted?
Exposure time: 2 minutes -
Immunocytochemistry/ Immunofluorescence - Anti-TDRD9 antibody (ab118427)ICC/IF image of ab118427 stained Hek293 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab118427, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) HeLa, HepG2 and MCF7 cells at 5µg/ml, and in 4% paraformaldehyde fixed (10 min) HeLa, Hek293, HepG2 and MCF7 cells at 5µg/ml. -
Western blot - Anti-TDRD9 antibody (ab118427)All lanes : Anti-TDRD9 antibody (ab118427) at 1 µg/ml (Milk blocking - 3%)
Lane 1 : Testis (Rat) Tissue Lysate
Lane 2 : Testis (Rat) Tissue Lysate with Immunizing peptide at 1 µg/ml
Lysates/proteins at 25 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 155 kDa
Observed band size: 171 kDa why is the actual band size different from the predicted?
Additional bands at: 55 kDa, 60 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 3 minutes
Abcam recommends using milk as the blocking agent (3%).
Protocols
Datasheets and documents
References (3)
ab118427 has been referenced in 3 publications.
- Sari I et al. Effect of ovarian stimulation on the expression of piRNA pathway proteins. PLoS One 15:e0232629 (2020). PubMed: 32365144
- Rocha-da-Silva L et al. Expression of genome defence protein members in proliferating and quiescent rat male germ cells and the Nuage dynamics. PLoS One 14:e0217941 (2019). PubMed: 31181099
- Murat P et al. RNA G-quadruplexes at upstream open reading frames cause DHX36- and DHX9-dependent translation of human mRNAs. Genome Biol 19:229 (2018). PubMed: 30591072
Images
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Western blot - Anti-TDRD9 antibody (ab118427)Anti-TDRD9 antibody (ab118427) at 1 µg/ml (Milk blocking - 3%) + Testis (Mouse) Tissue Lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 155 kDa
Observed band size: 171 kDa why is the actual band size different from the predicted?
Additional bands at: 190 kDa, 50 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 8 minutes
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TDRD9 antibody (ab118427)
IHC image of TDRD9 staining in Human normal testis formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab118427, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
-
Western blot - Anti-TDRD9 antibody (ab118427)All lanes : Anti-TDRD9 antibody (ab118427) at 1 µg/ml
Lane 1 : MCF7 cells (human).
Lane 2 : MDA231 cells (human).
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 155 kDa
Observed band size: 170 kDa why is the actual band size different from the predicted?
Exposure time: 2 minutes
-
Immunocytochemistry/ Immunofluorescence - Anti-TDRD9 antibody (ab118427)ICC/IF image of ab118427 stained Hek293 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab118427, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) HeLa, HepG2 and MCF7 cells at 5µg/ml, and in 4% paraformaldehyde fixed (10 min) HeLa, Hek293, HepG2 and MCF7 cells at 5µg/ml.
-
Western blot - Anti-TDRD9 antibody (ab118427)All lanes : Anti-TDRD9 antibody (ab118427) at 1 µg/ml (Milk blocking - 3%)
Lane 1 : Testis (Rat) Tissue Lysate
Lane 2 : Testis (Rat) Tissue Lysate with Immunizing peptide at 1 µg/ml
Lysates/proteins at 25 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 155 kDa
Observed band size: 171 kDa why is the actual band size different from the predicted?
Additional bands at: 55 kDa, 60 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 3 minutes
Abcam recommends using milk as the blocking agent (3%).